Alternative Hapten Design for Zearalenone Immunoreagent Generation
Appropriate hapten design and synthesis have been identified as critical steps to generate high-performance immunoreagents and to develop sensitive and selective immunoanalytical methods. Antibodies and immunoassays for the major mycotoxin zearalenone have been reported and marketed. However, zearal...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-03-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/14/3/185 |
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| author | Antonio Abad-Fuentes Consuelo Agulló Daniel López-Puertollano Ismael Navarro-Fuertes Antonio Abad-Somovilla Josep Vicent Mercader |
| author_facet | Antonio Abad-Fuentes Consuelo Agulló Daniel López-Puertollano Ismael Navarro-Fuertes Antonio Abad-Somovilla Josep Vicent Mercader |
| author_sort | Antonio Abad-Fuentes |
| collection | DOAJ |
| description | Appropriate hapten design and synthesis have been identified as critical steps to generate high-performance immunoreagents and to develop sensitive and selective immunoanalytical methods. Antibodies and immunoassays for the major mycotoxin zearalenone have been reported and marketed. However, zearalenone haptens have mostly been prepared by the oxime active ester technique, and hapten characterization has generally been poor or non-existent. In the present study, novel haptens of zearalenone with longer linkers and with alternative tethering sites have been designed for immunizing and assay conjugate preparation. All of these molecules were purified and spectroscopically verified, and a structure-activity relationship evaluation was carried out. This approach revealed that the hapten with the linker at the carbonyl group generated antibodies with a higher affinity than the hapten functionalized at the phenyl moiety. Antibodies produced with the latter hapten, on the other hand, showed lower cross-reactivity values to the major zearalenone metabolites. Finally, similar immunoassay sensitivity was achieved with all of the antibodies when heterologous haptens were employed. Furthermore, by altering the structure of the competing antigen, the immunoassay selectivity was modified. These results demonstrate that immunochemical methods for zearalenone rapid analysis can still be improved in terms of sensitivity and selectivity. |
| first_indexed | 2024-03-09T12:21:35Z |
| format | Article |
| id | doaj.art-2e6418dc7de24a86a21a7e42f2e4fb24 |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-03-09T12:21:35Z |
| publishDate | 2022-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-2e6418dc7de24a86a21a7e42f2e4fb242023-11-30T22:39:12ZengMDPI AGToxins2072-66512022-03-0114318510.3390/toxins14030185Alternative Hapten Design for Zearalenone Immunoreagent GenerationAntonio Abad-Fuentes0Consuelo Agulló1Daniel López-Puertollano2Ismael Navarro-Fuertes3Antonio Abad-Somovilla4Josep Vicent Mercader5Instituto de Agroquímica y Tecnología de Alimentos, Spanish Council for Scientific Research (CSIC), 28006 Madrid, SpainDepartment of Organic Chemistry, University of Valencia, 46010 València, SpainDepartment of Organic Chemistry, University of Valencia, 46010 València, SpainDepartment of Organic Chemistry, University of Valencia, 46010 València, SpainDepartment of Organic Chemistry, University of Valencia, 46010 València, SpainInstituto de Agroquímica y Tecnología de Alimentos, Spanish Council for Scientific Research (CSIC), 28006 Madrid, SpainAppropriate hapten design and synthesis have been identified as critical steps to generate high-performance immunoreagents and to develop sensitive and selective immunoanalytical methods. Antibodies and immunoassays for the major mycotoxin zearalenone have been reported and marketed. However, zearalenone haptens have mostly been prepared by the oxime active ester technique, and hapten characterization has generally been poor or non-existent. In the present study, novel haptens of zearalenone with longer linkers and with alternative tethering sites have been designed for immunizing and assay conjugate preparation. All of these molecules were purified and spectroscopically verified, and a structure-activity relationship evaluation was carried out. This approach revealed that the hapten with the linker at the carbonyl group generated antibodies with a higher affinity than the hapten functionalized at the phenyl moiety. Antibodies produced with the latter hapten, on the other hand, showed lower cross-reactivity values to the major zearalenone metabolites. Finally, similar immunoassay sensitivity was achieved with all of the antibodies when heterologous haptens were employed. Furthermore, by altering the structure of the competing antigen, the immunoassay selectivity was modified. These results demonstrate that immunochemical methods for zearalenone rapid analysis can still be improved in terms of sensitivity and selectivity.https://www.mdpi.com/2072-6651/14/3/185mycotoxinhapten designspacer armantibody affinityantibody specificityimmunoassay |
| spellingShingle | Antonio Abad-Fuentes Consuelo Agulló Daniel López-Puertollano Ismael Navarro-Fuertes Antonio Abad-Somovilla Josep Vicent Mercader Alternative Hapten Design for Zearalenone Immunoreagent Generation Toxins mycotoxin hapten design spacer arm antibody affinity antibody specificity immunoassay |
| title | Alternative Hapten Design for Zearalenone Immunoreagent Generation |
| title_full | Alternative Hapten Design for Zearalenone Immunoreagent Generation |
| title_fullStr | Alternative Hapten Design for Zearalenone Immunoreagent Generation |
| title_full_unstemmed | Alternative Hapten Design for Zearalenone Immunoreagent Generation |
| title_short | Alternative Hapten Design for Zearalenone Immunoreagent Generation |
| title_sort | alternative hapten design for zearalenone immunoreagent generation |
| topic | mycotoxin hapten design spacer arm antibody affinity antibody specificity immunoassay |
| url | https://www.mdpi.com/2072-6651/14/3/185 |
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